The purpose of this research is to identify metabolic pathways affected by intake of dietary bioactives and to determine their mechanisms of action. Bioactives of particular interest include polyphenols from berries, lycopene, beta-carotene and vitamin A, and isothiocyanates. Samples from previously conducted human studies will be used to support the objectives for the 2019 research activities. This Inter-Agency Agreement will capitalize on joint expertise of the NCI DCP NSRG and the USDA Beltsville Human Nutrition Research Center to conduct analyses on samples from human feeding interventions to better understand the role of diet in cancer prevention. More specifically, objectives of this research are: 1) to evaluate the metabolomics profile in urine and plasma using both NMR and LC-MS from the beta-carotene and lycopene tomato juice human feeding study conducted previously, 2) to analyze carotenoid absorption after consumption of freeze-dried broccoli, 3) to analyze isothiocyanates in urine after consumption of freeze dried broccoli, 4) to evaluate pro-inflammatory cytokines in blood after consumption of brassica vegetables, and 5) to investigate novel circulating biomarkers such as extracellular vesicles and their cargos and whether diet influences their abundance/composition.